title: Expression of rat histone H1d in Escherichia coli and its purification.
abstract: Histone H1 is involved in the folding of linear polynucleosomal filament into a 30-nm fiber. In an effort to understand the role of different domains of histone H1 in chromatin folding, we have now expressed rat histone H1d in Escherichia coli using pTrc99A expression vector by providing a 6-His tag at the C-terminus to facilitate its purification. The expressed protein histone H1d was purified from the soluble extract of E. coli by employing Ni2+ NTA-agarose and heparin-agarose chromatography. The recombinant histone H1d was shown to be authentic by its N-terminal amino acid analysis, its secondary structural characteristics, and its ability to (a) condense DNA and (b) bind specifically to synthetic four-way junction DNA.
authors.affiliation: Department of Biochemistry, Indian Institute of Science, Bangalore, 560 012, India.
authors.forename: M M
authors.initials: MM
authors.lastname: Bharath
authors.forename: J R
authors.initials: JR
authors.lastname: Khadake
authors.forename: M R
authors.initials: MR
authors.lastname: Rao
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000737:chemistry / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D006639:Histidine / Q000737:chemistry; D006657:Histones / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D011485:Protein Binding; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification
chemical_list: D017931:DNA Primers; D006657:Histones; D011994:Recombinant Proteins; D006639:Histidine; D004247:DNA